Two days before the LPS intraperitoneal injection, place mice in the anesthesia machine and once the mice are anesthetized shave abdominal area with a trimmer, as fur quenches the luminescence signal (Figure 1).

Figure 1. A representative picture of mice under anesthesia getting their abdominal area shaved with a trimmer

Weigh mice.

In the day of the experiment dilute LPS in DPBS and prepare syringes for injection.

Intraperitoneally inject mice with 2.5 mg/kg of LPS or the same volume DPBS for the control group (injection volume approximately 200 μl).

Figure 2. A representative example of in vivo imaging in 8 wk of age male C57BL/6 mice after i.p. injection of PBS (left) or LPS (2.5 mg/kg body weight) (right)

Figure 3. Screen capture image of the IVIS acquisition control panel

Quantify the MPO signal using the Living Image software. First select region of interest (ROI) using ROI tools and choose to automatically draw measurement ROIs and perform ROI analyses to measure photon radiance. Also measure background ROI and subtract from your ROI measurement. Use average radiance to plot your graph.

This protocol was adapted from a previously published study (de Almeida et al., 2015). This work was supported by grants from the National Institutes of Health (AI099009 and AR064349 to C.S., AR066739 to A.D., AI120625 and AI120618 to C.S. and A.D., T32AR007611 to L.d.A., and the American Heart Association 13GRNT17110117 to C.S.).

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